Our laboratory recently published a report demonstrating the ability of integration defective lentiviral vectors (IDLV's) to cure hemophilia B in preclinical settings, providing additional proof of their therapeutic potential. However, further improvements of the IDLV gene delivery system are required in order to establish IDLV's as a therapeutic modality for nonfatal human diseases such as hemophilia B. Thus, we propose: a) to employ a novel PPT-deleted vector to further reduce the low risk of insertional mutagenesis associated with systemic administration of IDLV's, b) to develop and test a novel IDLV-based human factor IX (hFIX) expression cassette as a means to minimize IDLV vector load required to fully correct FIX deficiency in hemophilia B mice, and c) to establish a novel packaging cell line to facilitate production of mobilization resistant IDLV's carrying expression cassettes in opposite orientation to the vector's LTR's. The proposal comprises three specific aims.
In Aim 1 we will focus on characterizing the ability of novel PPT-deleted IDLV's with reduced illegitimate integration to mediate efficient hepatic gene delivery, in vivo.
Aim 2 will focus on the development and testing of a novel hFIX cDNA with prolonged in vivo half-life (t1/2) and enhanced specific activity using IDLV's for hepatic gene delivery. The efficacy of the new IDLV's to correct FIX deficiency will be tested in hemophilia B mice. The focus of Aim 3 will be establishing of a novel RNA-regulated protein kinase (PKR) resistant packaging cell line generating high titers of gp64- pseudotyped vectors. A PPT-deletion will render the cell line-generated IDLV's less likely to illegitimately integrate, and internal expression cassettes incorporated in opposite orientation to the LTR's will render them less likely to be mobilized. Overall the proposed studies will yield a highly efficient and significantly safer gene delivery system most suitable for gene replacement therapy of nonfatal human diseases.
Here we propose to employ two mechanistically independent approaches to minimize the levels of illegitimate integration in IDLV's. We will employ novel PPT-deleted IDLV's, which exhibit lower levels of illegitimate integration, and will develop a highly efficacious hFIX. In addition, we propose to establish a novel packaging cell line to produce high titer IDLV's carrying expression cassettes in opposite orientation to the LTR's.
|Simmons, Aaron B; Bretz, Colin A; Wang, Haibo et al. (2018) Gene therapy knockdown of VEGFR2 in retinal endothelial cells to treat retinopathy. Angiogenesis 21:751-764|
|Becker, Silke; Wang, Haibo; Simmons, Aaron B et al. (2018) Targeted Knockdown of Overexpressed VEGFA or VEGF164 in Müller cells maintains retinal function by triggering different signaling mechanisms. Sci Rep 8:2003|
|Suwanmanee, Thipparat; Ferris, Martin T; Hu, Peirong et al. (2017) Toward Personalized Gene Therapy: Characterizing the Host Genetic Control of Lentiviral-Vector-Mediated Hepatic Gene Delivery. Mol Ther Methods Clin Dev 5:83-92|
|Hu, Peirong; Li, Yedda; Nikolaishvili-Feinberg, Nana et al. (2016) Hematopoietic Stem cell transplantation and lentiviral vector-based gene therapy for Krabbe's disease: Present convictions and future prospects. J Neurosci Res 94:1152-68|
|Hu, Peirong; Li, Yedda; Sands, Mark S et al. (2015) Generation of a stable packaging cell line producing high-titer PPT-deleted integration-deficient lentiviral vectors. Mol Ther Methods Clin Dev 2:15025|
|Monahan, Paul E; Sun, Junjiang; Gui, Tong et al. (2015) Employing a gain-of-function factor IX variant R338L to advance the efficacy and safety of hemophilia B human gene therapy: preclinical evaluation supporting an ongoing adeno-associated virus clinical trial. Hum Gene Ther 26:69-81|
|Shen, Shen; Berry, Garrett E; Castellanos Rivera, Ruth M et al. (2015) Functional analysis of the putative integrin recognition motif on adeno-associated virus 9. J Biol Chem 290:1496-504|
|Wang, Haibo; Yang, Zhihong; Jiang, Yanchao et al. (2014) Quantitative analyses of retinal vascular area and density after different methods to reduce VEGF in a rat model of retinopathy of prematurity. Invest Ophthalmol Vis Sci 55:737-44|
|Jiang, Yanchao; Wang, Haibo; Culp, David et al. (2014) Targeting Müller cell-derived VEGF164 to reduce intravitreal neovascularization in the rat model of retinopathy of prematurity. Invest Ophthalmol Vis Sci 55:824-31|
|Suwanmanee, Thipparat; Hu, Genlin; Gui, Tong et al. (2014) Integration-deficient lentiviral vectors expressing codon-optimized R338L human FIX restore normal hemostasis in Hemophilia B mice. Mol Ther 22:567-574|
Showing the most recent 10 out of 23 publications